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Lysophosphatidylethanolamine Is a Substrate for the Short-Chain Alcohol Dehydrogenase SocA from Myxococcus xanthus ^,

Department of Microbiology, University of Georgia, Athens, Georgia 30602,¹ Center for Biomarker Analysis and Department of Microbiology, University of Tennessee, Knoxville, Tennessee 37932-2575,² Department of Environmental Microbiology, UFZ Leipzig-Halle, D-04318 Leipzig, Germany³

Received 18 July 2006/ Accepted 29 September 2006

Short-chain alcohol dehydrogenases (SCADHs) synthesize a variety of intercellular signals and other chemically diverse products. It is difficult to predict the substrate of a SCADH on the basis of amino acid sequence homology, as the substrates are not known for most SCADHs. In Myxococcus xanthus, the SCADH CsgA is responsible for C signaling during fruiting body development, although the mechanism is unclear. Overexpression of the SCADH SocA compensates for the lack of CsgA and restores development and C signaling in csgA mutants. The potential of SocA in generating the C signal enzymatically was explored by developing a dehydrogenase assay-based screen to purify the SocA substrate(s). A SocA substrate was extracted from M. xanthus cells with acidified ethyl acetate and sequentially purified by solid-phase extraction on silica gel and by reverse-phase high-performance liquid chromatography. The fraction with the highest SocA dehydrogenase activity contained the lysophospholipid 1-acyl 2-hydroxy-sn-glycerophosphoethanolamine (lyso-PE) as indicated by the fragment ions and a phosphatidylethanolamine-specific neutral loss scan following liquid chromatography coupled to mass spectrometry. The abundant lysophospholipid with the mass m/z 450 (molecular ion [M-H]^–) had a monounsaturated acyl chain with 16 carbons. SocA oxidizes lyso-PE containing either saturated or unsaturated fatty acids but exhibits poor activity on L--glycerophosphorylethanolamine, suggesting that an acyl chain is important for activity. Of the five different head groups, only ethanolamine showed appreciable activity. The apparent K_m and V_max for lyso-PE 18:1 were 116 µM and 875 µmol min^–1 mg^–1, respectively. The catalytic efficiency (k_cat/K_m) was 1 x 10⁸ M^–1 s^–1. The proposed product, 1-acyloxy-3-(2-aminoethylphosphatyl) acetone was unstable, and the fragmented products were unable to rescue csgA mutant development. The active fraction from thin-layer chromatography also contained an unidentified SocA substrate that had morphogenic properties.

Published ahead of print on 6 October 2006.

Dedicated to David C. White, 1929-2006.

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